Project Summary Genetically dominant mutations in the gene that encodes peripheral myelin protein 22 (PMP22) lead to single amino acid changes in its sequence that result in defective myelin, underlying the common human peripheral neuropathy, Charcot-Marie-Tooth Disease Type IA (CMTD). It is believed that CMTD mutations result in misassembly of PMP22 early in the secretory pathway, resulting in the loss of protein function and also in the formation of potentially cytotoxic aggregates. The overall goal of this project is to elucidate the molecular biophysical nature of the perturbations made by CMTD-associated mutations to the structure, stability and folding of this critical membrane protein. We also seek to test whether chemical chaperones can correct the folding defects normally observed for CMTD mutant forms of PMP22. Aim 1. Characterize the structures of the wild type (WT) and CMTD mutant forms of human PMP22 using NMR spectroscopy. Aim 1 will test the hypothesis that CMTD mutant forms of PMP22 differ from the WT protein in terms of conformation and/or oligomeric state. Structural information will also illuminate PMP22's structure/function relationships and its role in myelin production and maintenance, and will provide biophysical insight into how amino acid mutations result in CMTD. Determination of PMP22's structure will also add to the currently sparse gallery of human membrane protein structures. Aim 2. Characterize the stability and folding kinetics of WT and CMTD mutant forms of PMP22. Aim 2 will test the hypothesis that disease-related mutations of PMP22 destabilize the protein. Aim 2 will also test the hypothesis that disease-related mutant forms of PMP22 fold more slowly and/or inefficiently than the wild type protein. We will also test whether CMTD mutant forms of PMP22 are aggregation-prone. Aim 3. Determine whether chemical chaperones can increase the cell surface expression of CMTD mutant forms of PMP22. Aim 3 will test the hypothesis that PMP22 is akin to other human membrane proteins that are linked to diseases involving protein misassembly and for which it has already been established that proper folding and trafficking can be restored using chemical chaperones.